During the hand-eye calibration process of the manipulator, the Euclidean distance error of the calibration results cannot be calculated because the true values of the hand-eye conversion matrix cannot be obtained. In this paper, a new method of projection error analysis is presented. Error analysis is carried out by using a priori knowledge that the location of AR markers is fixed in the calibration process. The coordinates of the AR marker center point are reprojected to the pixel coordinate system, and then compared with the true pixel coordinates of the AR marker center point obtained by corner detection or manual labeling in order to obtain the Euclidean distance between the two coordinates as the basis for error analysis. Experimental results show that the proposed optimization method can greatly improve the accuracy of hand-eye calibration results.